PT103582A - PRE-FERMENTED SYMBIOTIC MATRIX BASED ON A SUSPENSION OF OXYGEN AND ENCAPSULATED PROBIOTICS, PROCESS OF OBTAINING AND USING THEIR USE - Google Patents

Abstract

The present invention relates to a pre-fermented symbiotic matrix based on a suspension of cereals, preferably the oats, containing encased and prebiotic probotics, the process for obtaining and using them. OBJECT OF THE PRESENT INVENTION THE DEVELOPMENT OF A SYMBIOTIC MATRIX OF CEREALS, PREFERENTIALLY THE OATMEAL, PREFERENTIAL WITH FREE AND / OR ENCAPSULATED ENCAPSULATED PREBIOTIC COMPOUNDS, IN ORDER TO COMPLEMENT THE CURRENT FUNCTIONAL FOOD MARKET AND TO SOLVE THE PROBLEMS INHERENT TO THE REDUCED TIME OF THE VALIDITY PERIOD FOR THE LOSS OF VIABILITY OF THE PROBIOTICS TO VALUES BELOW THE MINIMUM LIMITS NECESSARY FOR THE PROMOTION OF THE BIOLOGICAL ACTIVITY. In addition to the foregoing, it is the object of the present invention to improve the conditions of the fermentative processes at different levels such as reducing the fermentation time, thus improving the energy required for the process, reducing the risk of contamination and maintaining microbial stability over the long term. THE PRE-FERMENTED SYMBIOTIC MATRIX IS INTENDED, IN PARTICULAR IN CASES IN WHICH THE INTOLERANCE AND / OR ALLERGY IN THE INTAKE OF MILK PRODUCTS IS VERIFIED, BEING STILL APPLICABLE TO THE PHARMACEUTICAL, COSMETIC AND PREFERENTIALLY FOOD INDUSTRY, INCLUDING ANIMAL FOOD.

Description

1

DESCRIPTION

PRE-FERMENTED SYMBIOTIC MATRIX BASED ON SUSPENSION OF ENCAPSULATED CEREALS AND PROBIOTICS, OBTAINED PROCESS AND USEFUL USE "

Field of the Invention The present invention relates to the preparation of a pre-fermented symbiotic matrix containing prebiotic and / or probiotic compounds, is applicable to the pharmaceutical, cosmetic and preferably food industry, including animal feed. The pre-fermented symbiotic matrix, free or not of dairy components, applies to the entire population, particularly in cases where intolerance and / or allergy to dairy products is present.

SUMMARY The object of the present invention is the development of a symbiotic matrix of cereals, preferably oats, pre-fermented with encapsulated probiotic and free and / or encapsulated probiotic compounds, with the aim of complementing the current functional food market and solving inherent problems to the shortest period of validity. In addition to the foregoing, it is an object of the present invention to improve the conditions of the fermentative processes at different levels, such as reducing the fermentation time, making the energy necessary for the process, reducing the risk of contamination and maintaining long-term microbial stability.

Background of the Invention 2

Over the last few decades, knowledge about the influence of diet on human health has been rising dramatically, with populations around the world becoming aware of the need for "healthy diets", as a matter of public health, which becomes present as the average life expectancy increases. With the growing popularity of functional foods among consumers, food companies have been faced with the challenge of manufacturing such foods in order to respond appropriately to market demands. All foods are functional in the sense of the term, as they provide the energy and nutrients needed for survival. A food can be considered as functional when there is a demonstration of a proven beneficial effect, in addition to nutritional effects, and should be able to improve the health and well-being of the consumer.

This market is characterized by being dynamic and innovative with a market share of 10 to 15% and a growth rate of 20 to 30% per year worldwide.

Probiotics can generally be defined as viable microorganisms, which beneficially affect the host because they promote the balance of their intestinal bacterial ecosystem. Lactobacillus, Bifidobacterium and Enterococcus, genera considered to be potentially probiotic, do indeed provide host protection against infections - since they prevent the adherence, replication and / or virulence of specific enteropathogens. These probiotics also have a beneficial effect in controlling diarrhea as well as in reducing the risk of development of some forms of cancer. 3

Effects on blood cholesterol reduction are also described. Another possible and scientifically validated effect is the effect on the hydrolysis of lactose through the production of lactase (β-galactosidase) - which facilitates the digestion of this sugar, thus offering solutions for individuals intolerant to lactose. The beneficial effect of probiotic products is ensured when they contain a minimum of 106 CFU / ml, which is based on the assumption that the recommended daily allowance is 108 - 109 viable cells, achievable by ingestion of 100 g of fermented product containing 106 - 107 viable cells / ml. Probiotics naturally have limitations in their health benefits due to their sensitivity to certain technological and functional factors, for example high oxygen levels, acidic environments, freezing and the transit itself through the gastrointestinal tract. Encapsulation methods have therefore been applied in order to increase their survival rate by protecting such microorganisms from such adverse conditions. Microencapsulation is defined as " packaging technology " solids, liquids or gases in very small capsules, capable of releasing their contents at controlled rates under specific conditions. There are a number of microencapsulation techniques, including emulsion and spray drying. Emulsion encapsulation consists of adding a small volume of a solution containing microbial cells and polymers (discontinuous phase) to a large volume of vegetable oil (continuous phase). The mixture is then homogenized to form a water-in-oil emulsion. Once this emulsion is obtained, the water-soluble polymer should be insolubilized 4 through a saline solution in order to form small gel particles in the oil phase. The size of the capsules can be controlled by varying the speed and type of agitation, and the diameter of the saline addition mechanism. The emulsion encapsulation process is easily reproduced on an industrial scale with high levels of microorganism survival (80 to 95%). The resulting capsules have a wide range of sizes, having as an example 25 to 50 mm.

In turn, the atomization encapsulation method consists in drying an aqueous mixture of encapsulating agent with viable microbial cells using an atomizer. Drying occurs when the solution after being vaporized comes into contact with a stream of hot air (inlet temperature), and is subsequently, with the aid of a vacuum cleaner, collected in the respective container. This technology has as great advantages its low processing cost, ease of operation, the possibility of using thermosensitive functional ingredients, the high quality / stability of the capsules obtained and the easy production of large quantities. The diameter of the capsules obtained may range from 5 to 75 Âμm.

A prebiotic is by definition an nondigestible food ingredient that beneficially affects the host by selectively stimulating the growth and / or activity of one or a limited number of bacteria in the colon. The term symbiotic refers to the synergetic association of probiotic and prebiotic agents with physiological activity in the same food. 5

Prebiotics and their combination with probiotics currently consist of an investment in the encapsulated food industry in order to maintain their long-term stability and optimize the nutritional qualities of the associated product. US2001 / 0016220 discloses with components of encapsulated and biologically active food products, as well as process for their production and use. The components referred to in this document encompass plant fibers, including those from oats, soluble and insoluble polysaccharides in water, pectins, lignins and gums. The biologically active components encapsulated in the aforementioned plant fibers can be probiotic microorganisms, prebiotics, enzymes, nutrients, secondary or natural plant constituents, substances with antioxidant activity, etc. Encapsulation substances can be constituted by polysaccharides vegetable or microbial) emulsifiers, peptides, proteins and prebiotic substances.

As regards the process for obtaining these compounds, the document in question provides for the introduction of the biologically active compounds into a medium containing capsule-forming substances. Although this document describes cereal products containing biologically active compounds, such as probiotics and / or prebiotics, encapsulated in a matrix formed by the vegetable fibers of cereals, differs considerably from the invention in question since it describes an aqueous suspension of pre- fermentation, subsequent encapsulation of microorganisms, by emulsion, in fluidized bed or by drying, and subsequent addition of prebiotic compounds, that is, while in the document all prebiotic or probiotic compounds are encapsulated and are introduced into the matrix in a In the same phase, the invention in question describes a process of obtaining said compounds in several steps, with protection of the microbial activity through encapsulation of the microorganisms. WO2005.002367 discloses oat-based therapeutic products and compositions free from probiotic microorganisms comprising proteins, protein hydrolysates and emulsifying lipids. In addition, the products and compositions may further comprise β-glucans, plant sterols. The respective production process is carried out by enzymatic treatment of the oat fraction, for removal of the carbohydrates, preferably by hydrolysis. WO02.065855 discloses non-dairy products, based on cereal dispersions, containing β-glucans, proteins, natural sugars and proteins. The process for their preparation uses enzymes, particularly hydrolases, but also isomerases, applied to cereal suspensions. WO02.37984 discloses fermented products by microbial cultures based on suspensions of oats, soy and milk-free, and the process for their preparation. This document provides for the use of Lactobacillus and Streptococcus strains in the fermentation of oat suspension, as well as the inclusion of various compounds such as calcium hydrogen phosphate and / or calcium phosphate, β-glucans, maltose, maltodextrin, proteins, etc. in an aqueous suspension of oats, which is subsequently incubated for fermentation. WOOO.65930 discloses cereal products, in particular oats, for use as feedstock in the food industry. The process of obtaining these products involves the preparation of a suspension of cereals, from bran, flakes or cereal flour. This suspension is subsequently homogenized, at a certain temperature and pressure, so as to obtain an emulsion. The emulsion can then be fermented by microorganisms such as Lactobacillus, Bifidobacterium, among others, acidified and finally pasteurized or ultrapasteurized, and may be further provided in powder form. CA2383021 discloses symbiotic compositions based on β-glucans from cereals, obtained from flours or cereal extracts inoculated with bacteria, for fermentation. The bacteria Lactobacillus, Streptococcus and / or Bifidobacteria are inoculated in aqueous suspensions of cereals treated with α-amylases, and a stabilizing agent is further added. W02004.037191 describes liquid or frozen symbiotic products derived from soy or dairy products composed of a mixture of probiotic components (such as Lactobacillus and Bifidobacteria) and prebiotics, the latter being composed of polymers, in particular, by inulin or an oligofructose. The process for their preparation provides for mixing the prebiotic and probiotic compounds in the liquid phase, fermentation of this mixture until a pH of 4.5 is obtained and subsequent stirring to obtain the final product. In this final step, it is also possible to include a percentage of carbon dioxide. The matter disclosed in these six documents differs substantially from the subject matter of the present invention, since in these the fermentation of oat suspensions is described by the addition of free microorganisms or enzymes, in a single step, to said suspension of oats or cereals, further compounds may be included, while the subject invention describes suspensions of cereals pre-fermented by immobilized microorganisms, to which encapsulated or encapsulated probiotic and encapsulated probiotic compounds are added thereafter.

In this way, and because the microorganisms are in the free form, they consequently reduce the shelf-life of the final consumer products and decrease the stability / viability of the microorganisms along the storage chain and gastrointestinal tract, compared to pre-fermented symbiotic matrix with encapsulated probiotics (40-60% increase), object of the present invention.

This solution solves the problems inherent to the reduced shelf life of the patents WO02.37984, WOOO.65930, CA2383021, W02004.037191 (40-60% increase over existing products with free microorganisms) thereof and maintenance of long-term microbial stability. The object of the invention is the development of dry extruded cereals with surface microorganisms and / or introduced therein and with soluble fiber sources having as an example of application breakfast cereals and foods for animals. The object of the present invention is to provide a symbiotic matrix of cereals, preferably oats, pre-fermented with encapsulated probiotics and free and / or encapsulated probiotics, when applied together, with a stabilizing effect of the microorganisms in the product and the passage of this through the gastro-intestinal tract. In addition to the aforementioned, the functional claim of cholesterol reduction associated with β-glucans as a source of soluble, non-digestible -prebiotic fibers is further object of this invention. The suspension of cereals, preferably of oats, is presented in fresh, freeze-dried and frozen form, adapted to the needs of the parties involved in the purchase process and, consequently, with numerous applications in the food industry. WO2004.070026 discloses processes for the immobilization of yeasts on gel beads of k-carrageenan or alginate continuously in brewing, for example by forming an emulsion between the non-aqueous continuous phase (vegetable oil) and the dispersed aqueous phase (yeast inoculated K-carrageenan), using static mixers. This matter differs from that disclosed in the present invention, since the immobilization process described, although referring to the formation of an emulsion between a vegetable oil and a polymer inoculated with a microorganism, differs from that proposed by the present invention, since the microorganism described in the concerned document 10 is a yeast whereas those of the present invention include all types of probiotic microorganisms. A phased procedure to produce symbiotic products from pre-fermented cereal suspensions with the addition of encapsulated probiotics and subsequent incorporation of prebiotic compounds into the cereal matrix leads to the development of a superior product, not only in the which relates to nutritional aspects related to the maintenance of microbial stability in the long term, but also in terms of storage time and availability to the consumer.

The present invention discloses a symbiotic matrix of cereals, preferably oats, pre-fermented with encapsulated and prebiotic probiotic compounds, the process for their production and their use in various applications, in particular in the food industry, but also in the pharmaceutical industry and others.

The products thus obtained have the same organoleptic characteristics as the products obtained by traditional fermentation processes.

When they include encapsulated microorganisms, they also have the advantage of increasing their viability / stability both over the storage period and during passage into the gastrointestinal tract after ingestion. 11

The matrices also have a longer shelf life of 40-60% compared to existing products on the market.

By using this technology a pre-fermented product is obtained with residual quantities of microorganisms and with the same organoleptic characteristics as a traditional fermented product, thus constituting an added value. The microbial cell immobilization technique confers advantages over free cell systems such as: (i) reduction of the fermentation time by 50 to 60%; (ii) increased stability and microbial metabolism; (iii) reduced risk of contamination; (iv) higher cell density; (v) constant quality of the associated product, for example, to a decrease in the risk of post-acidification in the product by the action of probiotics; (vi) improved use of the substrate and (vii) reuse of the cells for a long period of time due to constant cell regeneration. The process of obtaining these products discloses a way of improving the conditions of fermentative processes at different levels such as (i) reusing the immobilized cells continuously, (ii) reducing the fermentation time, thus making the energy necessary for the process ) reducing the risk of contamination and (vi) maintaining long-term microbial stability.

DETAILED DESCRIPTION OF THE INVENTION 1. Process for Obtaining the Oat Suspension 12 1.1. preparation of oat concentrate at 5 - 20 (w / w)% from the weighing of flakes, meal and / or meal and subsequent mixing in water; 1.2. heating the above mixture for 5 to 20 minutes at a temperature range of 80 to 110 ° C with continuous stirring; 1.3. milling the resulting preparation; 1.4. filtration of the suspension obtained; 1.5. cooling the temperature of the mixture to between 25 and 48 ° C. 2. Pre-fermentation process associated with cell encapsulation by fluidized bed emulsion 2.1. Pre-fermentation process The pre-fermentation process is carried out in a fluidized bed reactor with microorganisms immobilized in capsules obtained from step 2.1.1 to 2.4.3.

The capsules are introduced into a column with porosity less than the diameter thereof to allow interaction of matrix microorganisms within the pressurized reactor with constant and controlled bi-directional circulation of nitrogen. The microorganisms immobilized inside the column are reused fermentation after fermentation until their metabolic properties are lost. 2.1.1. Preparation of the cell culture 2.1.1.1. Preparation of the inoculum from frozen cultures and subsequent recovery through two consecutive passages in MRS broth supplemented with 0.05% (w / v) L-cysteine-HCl; 2.1.1.2. Inoculation of 1 to 20% (v / v) in 1000 ml of MRS broth (from Man Rogosa and Sharpe) supplemented with 0.05% (w / v) L-cysteine-HCl and further growth, as an example, for 24 h at 37 ° C, under anaerobic conditions for Lactobacillus acidophilus Ki, and 48 h at 37 ° C under anaerobic conditions for Bifidobacterium animalis Bo and Bbl2; 2.1.1.3. Centrifugation of the resulting cultures at 4000 rpm for 15 minutes at 4øC and subsequent washing with a solution of, for example, 0.9% (w / v) NaCl and concentrated in 100 ml thereof. 2.2. Preparation of polymer solution 2.2.1. Preparation of a solution of a polymer, for example κ-carrageenan of 1 to 5% (w / v), with constant stirring of from 1 to 4 hours at 60 to 80 ° C, followed by cooling to a temperature range of 35 to 45 ° C. 2.3. Preparation of the oil solution 2.3.1. Vegetable oil blend with at least one of the following compounds: 0.2% (v / v) Tween 80 and / or a protective agent, non-limiting example, 0.5% (v / v) sodium. 2.4. Formation of capsules 2.4.1. Mixing of 1 to 20% (v / v) cell suspension (see 2.1) with the prepared 1 to 5% (m / v) polymer solution (see 2.3); 2.4.2. Add the resulting mixture to 75-98% of the prepared oil solution (see 2.3). The obtained solution is homogenized to form a water-in-oil emulsion; 2.4.3. Capsule formation occurs after the addition of a solution of, for example, 10 mM KCl in a temperature range of 4 to 8øC to the mixture from the previous point. 2.5. Operating conditions of the pre-fermentation process After obtaining the capsules with micro-organisms for use in a fluidized bed reactor, the pre-fermentation process is carried out at a temperature range of 37 to 48 ° C for 4 to 8 hours under conditions anaerobic (flow of circulating nitrogen) and sterile, resulting in a fermented matrix. This suspension is drained to the reactor where the incorporation of the remaining food ingredients takes place. 3. Process for the encapsulation of microorganisms by emulsification and / or spray drying The encapsulation of the microorganisms is performed using the encapsulation techniques: 3.1. Emulsion as described in point 2; 3.2. Atomization drying: 3.2.1. Preparation of a polymer cell suspension (see sections 2.1, 2.2 and 2.4.1); 3.2.2. Drying of 250 ml of the above mixture under the conditions of the inlet and outlet temperatures of 150-175 ° C and 50-80 ° C, respectively; 3.2.3. Addition of the resulting powder to the suspension of pre-fermented oats (2.5) in a proportion of 2 to 5% (m / v), to ensure 108-1010 CFU in the matrix. 4. Incorporation of food ingredients:

Mixing of ingredients into the matrix resulting from the previous process (3.2.3), with inulin as an example, in a concentration range of 1 to 3%, preservatives, as non-limiting example sea salt, among others. 5. Presentation Forms of the Matrix The prefermented symbiotic matrix based on a suspension of oats and encapsulated probiotics may be presented in fresh, lyophilized and / or frozen form. The fresh matrix may further be in gel form and / or extruded.

Lisbon, November 06, 2006

Claims (24)

A pre-fermented symbiotic matrix characterized in that it comprises a suspension of cereals with encapsulated microorganisms and at least one of the following compounds: Free and / or encapsulated prebiotics; • Free microorganisms; Other food ingredients. A pre-fermented symbiotic matrix according to claim 1, characterized in that the cereals may be in flakes, flour or meal. A pre-fermented symbiotic matrix according to claim 2, characterized in that the cereals are preferably oats. A pre-fermented symbiotic matrix according to claim 2, characterized in that the cereals are preferably oats combined with at least one other cereal and / or legumes usually applied in the food industry, such as non-limiting example barley and soybeans , respectively. A pre-fermented symbiotic matrix according to claims 3 and 4, characterized in that it comprises cereals and / or legumes which may come from organic farming and / or fair trade. A pre-fermented symbiotic matrix according to claim 1, characterized in that it comprises as a prebiotic source soluble β-glucan fibers in biologically active amounts. A pre-fermented symbiotic matrix according to the preceding claim, characterized in that it comprises as an additional source, soluble β-glucan fibers which may also be extracted from cereals and / or legumes. A pre-fermented symbiotic matrix according to claims 6 and 7, characterized in that soluble β-glucan fibers are preferably extracted from cereals / legumes. A pre-fermented symbiotic matrix according to claims 5 to 8, characterized in that it comprises at least 0.75% (w / w) of soluble β-glucan fibers. A pre-fermented symbiotic matrix according to Claims 6 to 9, characterized in that it comprises, in addition to the soluble β-glucan fibers, other prebiotic compounds, such as, for example, non-limiting inulin, fructooligosaccharides (FOS) and the chitosans. A pre-fermented symbiotic matrix according to claims 6 to 10, characterized in that it can comprise further food ingredients, which in addition to the prebiotic function may have further functions, such as for example non-limiting, organoleptic functions (having as non-limiting examples 3 flavorings and / or fruit pulps), texture (as enzymes example) and sweeteners. A pre-compounded symbiotic matrix according to claim 6 to 11, characterized in that it may further comprise additional sources of antioxidants, fatty acids, for example non-limiting, omega 3, omega 6 and derivatives thereof, vitamins and minerals for in addition to those in cereals and / or legumes, free or encapsulated. A pre-fermented symbiotic matrix according to claim 1, characterized in that the encapsulated probiotic and non-probiotic 'GRAS' microorganisms are preferably of the genera Bifidobacterium and Lactobacillus. A pre-fermented symbiotic matrix according to claim 13, characterized in that it comprises inoculated microorganisms in an amount of not less than 108 " IO10 CFU / g, to ensure that the final product, at the point of consumption, contains at least 106-108 CFU / g. A pre-fermented symbiotic matrix according to claim 13 to 14, characterized in that it comprises probiotic and non-probiotic 'GRAS' microorganisms, encapsulated with at least one of the following coating materials: proteins, polysaccharides, lipids and hydrocolloids. 4 A pre-fermented symbiotic matrix according to claim 15, characterized in that the capsules have macro, micro and / or nano dimensions. Process for obtaining a pre-fermented symbiotic matrix according to the previous claims, characterized in that it is carried out by unitary operations such as pre-fermentation, encapsulation and subsequent incorporation of food ingredients. Process for obtaining a pre-fermented symbiotic matrix according to claim 17, characterized in that the pre-fermentation is carried out by placing the cereal suspension in a reactor with microbial cells immobilized in macrocapsules, coated with proteins, polysaccharides , lipids and hydrocolloids. Process for obtaining a pre-fermented symbiotic matrix according to claim 17, characterized in that the encapsulation method is carried out, for example, by drying, atomizing, emulsifying or coacervating. A process for obtaining a pre-fermented symbiotic matrix according to claim 17, characterized in that the incorporation of the food ingredients is carried out after the pre-fermentation and encapsulation step. Process for obtaining a pre-fermented symbiotic matrix according to claims 17 to 20, characterized in that the final product is processed so that the symbiotic matrix can be presented in fresh, lyophilised form and / or frozen. Process for obtaining a pre-fermented symbiotic matrix according to the preceding claim, characterized in that the fresh matrix can still be in the form of a gel and / or extruded. Use of the pre-fermented symbiotic matrix according to the previous claims, characterized in that it is applied to the food, livestock and pharmaceutical / cosmetic industries. Use of the pre-fermented symbiotic matrix according to the preceding claim, characterized in that it is applied in the production of novel foods, with non-limiting examples of beverages, ice creams, and / or nutraceuticals. Lisbon, November 6, 2006